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Trastuzumab Antibody ELISA Kit

Reaktivität: Human, Maus, Ratte Colorimetric Bridging ELISA 31.25-125 ng/mL Plasma, Serum
Produktnummer ABIN4886400
  • Target Alle Trastuzumab Antibody Produkte
    Trastuzumab Antibody
    Reaktivität
    Human, Maus, Ratte
    Nachweismethode
    Colorimetric
    Methodentyp
    Bridging ELISA
    Detektionsbereich
    31.25-125 ng/mL
    Untere Nachweisgrenze
    31.25 ng/mL
    Applikation
    ELISA
    Verwendungszweck
    Quantification of antibodies to Trastuzumab
    Proben
    Plasma, Serum
    Analytische Methode
    Quantitative
    Spezifität
    Anti-Trastuzumab antibodies
    Produktmerkmale
    This immunogenicity assay employs the bridging ELISA technique.
    Bestandteile
    Coated microtiter plate, 96 wells
    QC samples - 4x50ul
    10X wash buffer - 25ml
    Assay buffer - 50ml
    1000X secondary antibody - 17ul
    1000X detection reagent - 17ul
    TMB - 12ml
    TMB stop solution - 12ml
    Plate sealers - 3
    Benötigtes Material
    Precision pipettes calibrated to deliver 5-1000μL
    Multi-channel pipette calibrated to deliver 50-200μL
    Plate shaker
    Disposable tips
    Vortex-Mixer
    Distilled or de-ionized water
    Microplate reader capable of reading 450nm with background subtrac
  • Applikationshinweise
    Optimal working dilution should be determined by the investigator.
    Probenmenge
    15 μL
    Testdauer
    3.5 h
    Plattentyp
    Pre-coated
    Protokoll
    The Trastuzumab immunogenicity assay employs the bridging ELISA technique. A precoated 96 well capture antibody plate is provided. Quality control and test samples are pipetted into the appropriate wells. Anti-Trastuzumab present in biological matrices binds the immobilized capture antibody. After washing away any unbound substances, secondary antibody is added to the wells and after a final wash a detection reagent is added. The plate is washed to remove any unbound antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of anti-Trastuzumab present in test samples. Four levels of QC samples give a qualitative reference signal which can be used to determine the level (High, Medium, Low, Negative) of anti-Trastuzumab antibody in the unknown samples.
    Aufbereitung der Reagenzien

    Prepare only the appropriate amount of required reagent on the day of use. Store all reagents as per instructions stated on the label. 1. Wash Buffer (1X) Preparation: Dilute wash buffer concentrate with ultra-pure water 1/10 before use (for example add 2 mL concentrate to 18 mL ultra-pure water). Mix well. 2. Secondary antibody (1X) Preparation: Dilute secondary antibody with assay buffer 1/1000 before use (for examples add 12 μL concentrate to 12 mL of assay buffer). Mix well. 3. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 12 μL concentrate to 12 mL of assay buffer). Mix well.

    Probennahme
    This kit is compatible with EDTA-plasma, heparinplasma and serum samples. Samples can be stored at or below -20 °C for up to 1 year.
    Aufbereitung der Proben

    Dilute QC samples and test samples 1/10 with assay buffer (for example add 30μL of prepared calibrator or sample to 270μL of assay buffer). Mix well. Do not store diluted samples. If test samples are out of range, then they may be further diluted.

    Testdurchführung

    This immunogenicity assay employs the bridging ELISA technique. Capture antibody is precoated onto a 96 well microplate. Quality control and test samples are pipetted into the appropriate wells. AntiTrastuzumab present in biological matrices is bound by the immobilized capture antibody. After washing away any unbound substances, secondary antibody is added to the wells and after a final wash a detection reagent is added. The plate is washed to remove any unbound antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of anti-Trastuzumab present in test samples. Three levels of QC samples give a qualitative reference signal which can be used to determine the level of anti-Trastuzumab antibody in the unknown samples. The color development is stopped and the intensity of the color is measured.

    Ergebnisberechnung
    1. Because anti-drug antibodies will vary in terms of affinity and concentration, this assay provides a qualitative readout. As such the user should use the comparable positive controls when comparing interassay results. The provided controls are tested for comparability between lots and can be traced. 2. The anti-drug antibody titers in the test samples will fall in the range of high, medium, low or negative. We recommend each lab develop their own statistical cutpoint using methodologies as described by G. Shankar, et al. (2008). (Recommendations for the validation of immunoassays used for detection of host antibodies against biotechnology products. J. Pharmaceutical and Biomedical Analysis 48:1267-1281). 3. Any sample undiluted or diluted still reading greater than the highest standard should be diluted appropriately with assay buffer and retested. If the samples have been diluted, the concentration determined from the standard curve must be multiplied by the dilution factor.
    Testpräzision
    Intra-assay precision: < 10%
    Inter-assay precision: < 10%
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Konservierungsmittel
    Without preservative
    Vorsichtsmaßnahmen
    Read manual completely before beginning
    Lagerung
    -20 °C
    Informationen zur Lagerung
    Store kit components at -20°C unless specified otherwise. DO NOT USE past kit expiration date. Some vials contain a small amount of reagents. Spin tubes on pulse setting prior to opening.
    Haltbarkeit
    12 months
  • Target Alle Trastuzumab Antibody Produkte
    Trastuzumab Antibody
    Abstract
    Trastuzumab Antibody Produkte
    Substanzklasse
    Antibody
    Hintergrund
    Trastuzumab (Herceptin® ) is a humanized recombinant monoclonal antibody used for the treatment of primary breast cancers overexpressing human epidermal growth factor 2 (HER2). HER2 protein is overexpressed in 25-30 % of breast cancers.
    Gen-ID
    2064
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